Methods and materials for reducing amyloid beta levels within a mammal

ABSTRACT

This document provides methods and materials for reducing amyloid beta levels within a mammal (e.g. a mammal having Alzheimer&#39;s disease). For example, this document provides methods for using compositions containing a potato polysaccharide preparation to reduce one or more symptoms of Alzheimer&#39;s disease. In some cases, a composition containing a potato polysaccharide preparation provided herein can be used to increase binding, sequestration, and/or degradation of CNS-derived amyloid beta polypeptides, thereby inhibiting the formation of neurofibrillary plaques.

BACKGROUND 1. Technical Field

This document relates to methods and materials for reducing amyloid betalevels within a mammal having Alzheimer's disease. For example, thisdocument relates to using compositions containing a potatopolysaccharide preparation to reduce one or more symptoms of Alzheimer'sdisease. In some cases, this document relates to using compositionscontaining a potato polysaccharide preparation to degrade CNS-derivedamyloid beta polypeptides.

2. Background Information

Potatoes are starchy, edible tubers obtained from potato plants and forman integral part of much of the world's food supply. In fact, potatoesare the fourth largest food crop in the world. The main potato speciesworldwide is Solanum tuberosum.

SUMMARY

This document provides methods and materials for reducing amyloid betalevels within a mammal (e.g., a mammal having Alzheimer's disease). Forexample, this document provides methods for using compositionscontaining a potato polysaccharide preparation to reduce one or moresymptoms of Alzheimer's disease. In some cases, a composition containinga potato polysaccharide preparation provided herein can be used toincrease binding, sequestration, and/or degradation of CNS-derivedamyloid beta polypeptides. In some cases, a composition containing apotato polysaccharide preparation provided herein can be used degradeCNS-derived amyloid beta polypeptides, for example at peripheral sitesof action and/or to inhibit the formation of neurofibrillary plaques.

Having the ability to use a composition containing a potatopolysaccharide preparation described herein to reduce one or moresymptoms of Alzheimer's disease can provide clinicians and patients withan effective treatment regime for improving a patient's quality of life.

This document also provides compositions (e.g., nutritional supplementcompositions) that contain a potato polysaccharide preparation. Forexample, this document provides nutritional supplement compositionscontaining a potato polysaccharide preparation, methods for obtainingpotato polysaccharide preparations, methods for making nutritionalsupplement compositions containing a potato polysaccharide preparation,and methods for increasing or decreasing expression of polypeptidesinvolved with Alzheimer's disease.

In some cases, the compositions provided herein (e.g., nutritionalsupplement compositions and potato polysaccharide preparations providedherein) can be used to increase or decrease expression of polypeptidesinvolved with Alzheimer's disease and the metabolism of CNS-derivedamyloid beta polypeptides. For example, a composition containing apotato polysaccharide preparation provided herein or a potatopolysaccharide preparation provided herein can be used to increaseexpression of a low density lipoprotein receptor-related protein 1(LRP1) polypeptide, an amyloid beta (A4) precursor protein binding,family B member 1, (APBB1) polypeptide, an insulin degrading enzyme(IDE) polypeptide, a glutathione peroxidase 3 (GPX3) polypeptide, aglutathione peroxidase 4 (GPX4) polypeptide, or a combination thereof.In some cases, a composition containing a potato polysaccharidepreparation provided herein or a potato polysaccharide preparationprovided herein can be used to decrease expression of an insulin-likegrowth factor 1 (IGF1), a nitric oxide synthase 2, inducible (NOS2), ora combination thereof.

In some cases, the compositions provided herein (e.g., nutritionalsupplement compositions and potato polysaccharide preparations providedherein) can be used to increase or decrease expression of polypeptidesinvolved with binding, sequestration, and/or degradation of CNS-derivedamyloid beta polypeptides in adipose tissue. For example, a compositioncontaining a potato polysaccharide preparation provided herein or apotato polysaccharide preparation provided herein can be used toincrease expression of an LRP1 polypeptide, an APBB1 polypeptide, an IDEpolypeptide, a GPX3 polypeptide, or a combination thereof.

In some cases, the compositions provided herein (e.g., nutritionalsupplement compositions and potato polysaccharide preparations providedherein) can be used to increase or decrease expression of polypeptidesinvolved with binding, sequestration, and/or degradation of CNS-derivedamyloid beta polypeptides in blood. For example, a compositioncontaining a potato polysaccharide preparation provided herein or apotato polysaccharide preparation provided herein can be used toincrease expression of an APBB1 polypeptide, an IDE polypeptide, a GPX4polypeptide, or a combination thereof.

In some cases, the compositions provided herein (e.g., nutritionalsupplement compositions and potato polysaccharide preparations providedherein) can be used to increase or decrease expression of polypeptidesinvolved with oxidative stress and proinflammatory pathways. Forexample, a composition containing a potato polysaccharide preparationprovided herein or a potato polysaccharide preparation provided hereincan be used to decrease expression of an insulin-like growth factor 1(IGF1) polypeptide, a nitric oxide synthase 2, inducible (NOS2)polypeptide, or a combination thereof.

In general, one aspect of this document features a method for reducingamyloid beta levels within a mammal having Alzheimer's disease. Themethod comprises, or consist essentially of, administering to the mammala composition comprising a potato polysaccharide preparation obtainedfrom raw potatoes, wherein the level of amyloid beta within the mammalis reduced. The mammal can be a human. The level of amyloid beta withinthe mammal can be reduced in blood. The level of amyloid beta within themammal can be reduced in adipose tissue. The composition can comprisethe potato polysaccharide preparation in an amount that results inbetween 0.05 mg and 50 mg of the potato polysaccharide component of thepotato polysaccharide preparation being administered to the mammal perkg of body weight of the mammal. The composition can comprise between 1mg and 100 mg of the potato polysaccharide preparation. The compositioncan comprise between 6 mg and 20 mg of the potato polysaccharidepreparation. The composition can comprise between 1 mg and 100 mg of thepotato polysaccharide component of the potato polysaccharidepreparation. The composition can comprise between 6 mg and 20 mg of thepotato polysaccharide component of the potato polysaccharidepreparation. The composition can be in the form of a tablet. Thecomposition can comprise alpha lipoic acid. The composition can comprisealpha tocopherol. The potato polysaccharide preparation can be in anamount that results in between 0.075 mg and 0.5 mg of the potatopolysaccharide component of the potato polysaccharide preparation beingadministered to the mammal per kg of body weight of the mammal. At leastabout 80 percent of the potato polysaccharide preparation can be potatopolysaccharide. At least about 90 percent of the potato polysaccharidepreparation can be potato polysaccharide. At least about 95 percent ofthe potato polysaccharide preparation can be potato polysaccharide.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention pertains. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

Other features and advantages of the invention will be apparent from thefollowing detailed description, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is an HPLC chromatogram of a 10% ACN extract of raw potato(Russet Burbank).

FIG. 2 is an HPLC chromatogram of collected and re-purified 3.5 minutepeak material from a 10% ACN extract of raw potato shown in FIG. 1.

FIG. 3 is an LC/MS trace of 3.5 minute HPLC peak material.

FIG. 4 is a full NMR spectrum of 3.5 minute HPLC peak material.

FIG. 5 is an expanded NMR spectrum of 3.5 minute HPLC peak material.

FIG. 6 is a total ion chromatogram of derivatized carbohydrate fragmentsof 3.5 minute HPLC peak material obtained from raw potato RussetBurbank).

FIG. 7 is a fragmentation pattern of diacetamide. The peak fragmentationpattern is in the top panel, the compound library fragmentation match isin the bottom panel, and an overlay of the two is in the center panel.

FIG. 8 is a fragmentation pattern of 3-acetoxy pyridine. The peakfragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 9 is a fragmentation pattern of 3,4-furan dimethanol, diacetate.The peak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 10 is a fragmentation pattern of 1,2,3-propanetriol diacetate. Thepeak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 11 is a fragmentation pattern of imidazole, 2-acetamino-5-methyl.The peak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 12 is a fragmentation pattern of6,7-dihydro-5H-pyrrol[2,1,c][1,2,4] triazole-3-carboxylic acid. The peakfragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 13 is a fragmentation pattern of acetic acid,1-(2-methyltetrazol-5-yl) ethenyl ester. The peak fragmentation patternis in the top panel, the compound library fragmentation match is in thebottom panel, and an overlay of the two is in the center panel.

FIG. 14 is a fragmentation pattern of 1,2,3,4-butanetriol, tetraacetate(isomer 1). The peak fragmentation pattern is in the top panel, thecompound library fragmentation match is in the bottom panel, and anoverlay of the two is in the center panel.

FIG. 15 is a fragmentation pattern of 1,2,3,4-butanetriol, tetraacetate(isomer 2). The peak fragmentation pattern is in the top panel, thecompound library fragmentation match is in the bottom panel, and anoverlay of the two is in the center panel.

FIG. 16 is a fragmentation pattern of pentaerythritol tetraacetate. Thepeak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 17 is a fragmentation pattern of 1,2,3,4,5-penta-o-acetyl-D-xylitol(isomer 1). The peak fragmentation pattern is in the top panel, thecompound library fragmentation match is in the bottom panel, and anoverlay of the two is in the center panel.

FIG. 18 is a fragmentation pattern of 1,2,3,4,5-penta-o-acetyl-D-xylitol(isomer 2). The peak fragmentation pattern is in the top panel, thecompound library fragmentation match is in the bottom panel, and anoverlay of the two is in the center panel.

FIG. 19 is a fragmentation pattern of 3,5-diacetoxy benzyl alcohol. Thepeak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 20 is a fragmentation pattern of β-D-galactopyranose, pentaacetate.The peak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 21 is a fragmentation pattern of D-mannitol hexaacetate. The peakfragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 22 is a fragmentation pattern of galacticol, hexaacetate. The peakfragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 23 is a fragmentation pattern of cyclohexane carboxylic acid,1,2,4,5-tetrakis(acetoxy), (1α,3α,4α,5β)-(−). The peak fragmentationpattern is in the top panel, the compound library fragmentation match isin the bottom panel, and an overlay of the two is in the center panel.

FIG. 24 is a fragmentation pattern of muco-inositol, hexaacetate. Thepeak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 25 is a fragmentation pattern of D-glucitol-hexaacetate. The peakfragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 26 is a fragmentation pattern of myo-inositol, hexaacetate. Thepeak fragmentation pattern is in the top panel, the compound libraryfragmentation match is in the bottom panel, and an overlay of the two isin the center panel.

FIG. 27 is an HPLC chromatogram of a 10% ACN extract of raw OrganicYellow potato.

FIG. 28 is an HPLC chromatogram of a 10% ACN extract of raw Purplepotato.

FIG. 29 is an HPLC chromatogram of a 10% ACN extract of raw Idaho Russetpotato.

FIG. 30 is an HPLC chromatogram of a 10% ACN extract of raw Yukon Goldpotato.

FIG. 31 is an HPLC chromatogram of a 10% ACN extract of raw sweetpotato.

FIG. 32 is an HPLC chromatogram of a 10% ACN extract of boiled Purplepotato.

FIG. 33 is an HPLC chromatogram of two pooled fraction collections fromIdaho Russet potatoes.

FIG. 34 is an HPLC chromatogram of fractions collections from 3 g ofpurple potatoes.

FIG. 35 is a real time PCR amplification plot for IDE demonstratingdifferences in threshold cycle numbers between potato polysaccharidepreparation treated ZDF and untreated control ZDF in rat adipose tissuesamples. The higher cycle number for the control rat's tissue equates toa lower gene expression.

FIG. 36 is a Real time PCR amplification plot for IDE demonstratingdifferences in threshold cycle numbers between potato polysaccharidepreparation treated ZDF and untreated control ZDF in rat blood samples.The higher cycle number for the control rat's tissue equates to a lowergene expression.

FIG. 37 is a graph plotting percent change in IDE gene expression, asmeasured by real time PCR analyses. The enhancements in IDE geneexpression were determined to be 44.9±2.8% (n=9, p=0.02) and 51.3±6.2%(n=6, p=0.04), for adipose tissue and blood samples, respectively viaUnpaired t-tests.

DETAILED DESCRIPTION

This document provides methods and materials for reducing amyloid betalevels within a mammal (e.g., a mammal having Alzheimer's disease). Asdescribed herein, a composition containing a potato polysaccharidepreparation can be used to reduce one or more symptoms of Alzheimer'sdisease. For example, a composition containing a potato polysaccharidepreparation provided herein can be administered to any appropriatemammal (e.g., rat, mouse, dog, cat, horse, cow, goat, pig, chicken,duck, rabbit, sheep, monkey, or human) to reduce one or more symptoms ofAlzheimer's disease. Examples of Alzheimer's disease symptoms include,without limitation, difficulty in remembering recent events (e.g., shortterm memory loss), problems with language, disorientation (includingeasily getting lost), mood swings, loss of motivation, not managingself-care, behavioral issues, or combinations thereof.

Any appropriate route of administration (e.g., oral or parenteraladministration) can be used to administer a composition containing apotato polysaccharide preparation provided herein (e.g., a nutritionalsupplement composition provided herein) to a mammal. For example, acomposition containing a potato polysaccharide preparation providedherein can be administered orally. In some cases, a compositioncontaining a potato polysaccharide preparation provided herein can beadministered by injection.

A composition provided herein (e.g., a nutritional supplementcomposition) can include one or more potato polysaccharide preparations.A potato polysaccharide preparation can be a preparation that isobtained from a water extract of potato and that contains polysaccharidematerial having the ability to be eluted from a C18 cartridge (e.g., aSep-Pak Plus C-18 cartridge) with 10% acetonitrile. In some cases, apotato polysaccharide preparation can be a preparation that is obtainedfrom potato and that contains polysaccharide material having HPLCcharacteristics of that of the peak eluted at 3.5 minutes as describedin Example 1 (see, also, FIGS. 1, 2, and 27-33). In some cases, apolysaccharide of a potato polysaccharide preparation provided hereincan be a polar, water-soluble polysaccharide. In some cases, apolysaccharide of a potato polysaccharide preparation provided hereincan be a highly substituted complex xyloglucan material.

In some cases, a potato polysaccharide preparation can be a preparationthat is obtained from potato and that contains polysaccharide materialthat, when derivatized, results in at least the following acylatedcarbohydrates as assessed using GC/MS: (a) myo-inositol (set to 1× toserve as an internal standard), (b) glucose at about 40× to about 60×the myo-inositol content (e.g., glucose at about 50× the myo-inositolcontent), (c) xylose at about 10× to about 20× the myo-inositol content(e.g., xylose at about 15× the myo-inositol content), (d) mannose atabout 5× to about 15× the myo-inositol content (e.g., mannose at about10× the myo-inositol content), and (e) galactose at about 3× to about 7×the myo-inositol content (e.g., galactose at about 5× the myo-inositolcontent). The derivatization procedure can include forming a dry residueof the polysaccharide material that is then hydrolyzed usingtrifluoroacetic acid. The resulting material is then reduced usingsodium borohydride, and after borate removal, the end product isacylated using acetic anhydride and pyridine. The end products of thereaction are then injected directly on GC/MS to identify the acylatedcarbohydrates.

In some cases, a potato polysaccharide preparation can be a preparationthat is obtained from potato and that contains polysaccharide materialthat, when derivatized and assessed using GC/MS, results in at leastfour major components (3,4-furan dimethanol, diacetate;1,2,3,4,5-penta-o-acetyl-D-xylitol (isomer 1); 3,5-diacetoxy-benzylalcohol; and D-glucitol-hexaacetate). See, e.g., Example 1. In somecases, a potato polysaccharide preparation can be a preparation that isobtained from potato and that contains polysaccharide material that,when derivatized and assessed using GC/MS, results in the compoundslisted in Table 1 or results in the profile shown in FIG. 6.

In some cases, a potato polysaccharide preparation provided herein canbe a substantially pure potato polysaccharide preparation. Typically, asubstantially pure potato polysaccharide preparation is a preparationthat contains a single peak of material (e.g., a single peak ofpolysaccharide material) when assessed using, for example, HPLC (see,e.g., FIGS. 2 and 32). In some cases, greater than 60, 70, 75, 80, 85,90, 95, or 99 percent of a potato polysaccharide preparation providedherein can be polysaccharide material obtained from a potato.

Any appropriate potato species or variety can be used to obtain a potatopolysaccharide preparation provided herein. For example, Solanumtuberosum, Ipomoea batatas, S. acaule, S. bukasovii, S. leptophyes, S.megistacrolobum, S. commersonii, or S. infundibuliforme can be used toobtain a potato polysaccharide preparation provided herein. In somecases, potato varieties of S. tunerosum such as Organic Yellow, Purpleor blue varieties, Cream of the Crop, Adirondack Blue, Adirondack Red,Agata, Almond, Andes Gold, Andes Sun, Apline, Alturas, Amandine,Annabelle, Anya, Arran Victory, Atlantic, Avalanche, Bamberg, BannockRusset, Belle de Fontenay, BF-15, Bildtstar, Bintje, Blazer Russet, BlueCongo, Bonnotte, British Queens, Cabritas, Camota, Canela Russet, Cara,Carola, Chelina, Chiloé, Cielo, Clavela Blanca, Désirée, Estima, Fianna,Fingerling, Flava, German Butterball, Golden Wonder, Goldrush, HomeGuard, Innovator, Irish Cobbler, Jersey Royal, Kennebec, Kerr's Pink,Kestrel, Keuka Gold, King Edward, Kipfler, Lady Balfour, Langlade,Linda, Marcy, Marfona, Maris Piper, Marquis, Megachip, Monalisa, Nicola,Pachacoña, Pike, Pink Eye, Pink Fir Apple, Primura, Ranger Russet,Ratte, Record, Red LaSoda, Red Norland, Red Pontiac, Rooster, RussetBurbank, Russet Norkotah, Selma, Shepody, Sieglinde, Silverton Russet,Sirco, Snowden, Spunta, Up to date, Stobrawa, Superior, Vivaldi,Vitelotte, Yellow Finn, or Yukon Gold can be used to obtain a potatopolysaccharide preparation provided herein.

Any appropriate method can be used to obtain a potato polysaccharidepreparation provided herein. For example, raw potato material can behomogenized (e.g., homogenized with a Polytron homogenizer) in water andmaintained at room temperature for a period of time (e.g., about 1 hour)with occasional shaking. The homogenate can be centrifuged (e.g.,centrifuged at 4000 g for 10 minutes) to remove any larger solidmaterial. The resulting supernatant can be loaded onto a Solid PhaseExtraction cartridge (e.g., a C18 cartridge such as a Sep-Pak Plus C-18cartridge), and the polysaccharide material eluted with 10 percentacetonitrile. Once eluted, the polysaccharide material can be dried andstored (e.g., stored at about 4° C.).

This document also provides nutritional supplement compositionscontaining one or more potato polysaccharide preparations providedherein. For example, a potato polysaccharide preparation provided hereinobtained from Idaho Russet potatoes can be formulated into a nutritionalsupplement composition.

Any appropriate dose of a potato polysaccharide preparation providedherein can be used to formulate a composition provided herein (e.g., anutritional supplement composition or potato polysaccharide preparationprovided herein). For example, a potato polysaccharide preparationprovided herein can be used to formulate a composition for reducingamyloid beta levels within a mammal having Alzheimer's disease. Thecomposition can contain between about 1 mg and about 750 mg (e.g.,between about 1 mg and about 500 mg, between about 1 mg and about 250mg, between about 5 mg and about 40 mg, between about 5 mg and about 30mg, between about 5 mg and about 20 mg, between about 6 mg and about 50mg, between about 6 mg and about 20 mg, between about 10 mg and about 25mg, or between about 15 mg and about 20 mg) of the potato polysaccharidecomponent of the potato polysaccharide preparation. In some cases, acomposition (e.g., a nutritional supplement composition) can beformulated to deliver about 0.05 mg of the potato polysaccharidecomponent per kg of body weight to about 0.5 mg of the potatopolysaccharide component per kg of body weight to a mammal (e.g., ahuman) per day. For example, a nutritional supplement composition can beformulated into a single oral composition that a human can swallow oncea day to provide between about 0.05 mg of the potato polysaccharidecomponent per kg of body weight to about 0.5 mg of the potatopolysaccharide component per kg of body weight.

Any appropriate method can be used to formulate a composition providedherein (e.g., a nutritional supplement composition or potatopolysaccharide preparation provided herein). For example, commonformulation mixing techniques and preparation techniques can be used tomake a composition (e.g., a nutritional supplement composition) havingthe components described herein. In addition, a composition providedherein (e.g., a nutritional supplement composition or potatopolysaccharide preparation provided herein) can be in any form. Forexample, a composition provided herein (e.g., a nutritional supplementcomposition or potato polysaccharide preparation provided herein) can beformulated into a pill, capsule, tablet, gel cap, nutritional shake,nutritional bar, rectal suppository, sublingual suppository, nasalspray, inhalant, or injectable ampule. In some cases, a compositionprovided herein (e.g., a nutritional supplement composition) can includeone or more potato polysaccharide preparations provided herein alone orin combination with other ingredients including, without limitation,gelatin, cellulose, starch, sugar, bentonite, lactic acid, mannitol,alpha lipoic acid, alpha tocopherol, L-ascorbate, or combinationsthereof.

In some cases, a composition containing a potato polysaccharidepreparation provided herein or a potato polysaccharide preparationprovided herein can be used to increase expression of a LRP1polypeptide, an APBB1 polypeptide, an IDE polypeptide, a GPX3polypeptide, a GPX4 polypeptide, or a combination thereof. In somecases, a composition containing a potato polysaccharide preparationprovided herein or a potato polysaccharide preparation provided hereincan be used to decrease expression of an IGF1 polypeptide, a NOS2polypeptide, or a combination thereof.

In humans, a composition containing a potato polysaccharide preparationprovided herein or a potato polysaccharide preparation provided hereincan be used to increase expression of a human LRP1 polypeptide, a humanAPBB1 polypeptide, a human IDE polypeptide, a human GPX3 polypeptide, ahuman GPX4 polypeptide, or a combination thereof. In some cases, acomposition containing a potato polysaccharide preparation providedherein or a potato polysaccharide preparation provided herein can beused to decrease expression of a human IGF1 polypeptide, a human NOS2polypeptide, or a combination thereof.

A human LRP1 polypeptide can have the amino acid sequence set forth inGenBank® Accession No. NP_002323.2 (GI No. 126012562) and can be encodedby the nucleic acid sequence set forth in GenBank® Accession No.NG_016444 (GI No. 284813599). A human APBB1 polypeptide can have theamino acid sequence set forth in GenBank® Accession No. O00213.2 (GI No.12229629) and can be encoded by the nucleic acid sequence set forth inGenBank′ Accession No. NG_029615.1 (GI No. 342349296). A human IDEpolypeptide can have the amino acid sequence set forth in GenBank®Accession No. AAA52712.1 (GI No. 184556) and can be encoded by thenucleic acid sequence set forth in GenBank® Accession No. NG_013012.1(GI No. 260593646). A human GPX3 polypeptide can have the amino acidsequence set forth in GenBank® Accession No. NP_002075.2 (GI No.6006001) and can be encoded by the nucleic acid sequence set forth inGenBank® Accession No. NC_000005.10 (GI No. 568815593). A human GPX4polypeptide can have the amino acid sequence set forth in GenBank®Accession No. AAH22071.1 (GI No. 34784795) and can be encoded by thenucleic acid sequence set forth in GenBank® Accession No. NC_000019.10(GI No. 568815579). A human IGF1 polypeptide can have the amino acidsequence set forth in GenBank® Accession No. NP_001104753.1 (GI No.163659899) and can be encoded by the nucleic acid sequence set forth inGenBank® Accession No. NC_000012.12 (GI No. 568815586). A human NOS2polypeptide can have the amino acid sequence set forth in GenBank®Accession No. AAI30284.1 (GI No. 120660146) and can be encoded by thenucleic acid sequence set forth in GenBank® Accession No. NG_011470.1(GI No. 22480926).

In some cases, a potato polysaccharide preparation provided herein or anutritional supplement composition provided herein can be used toincrease expression of polypeptides involved with the metabolism ofCNS-derived amyloid beta polypeptides within the adipose tissuecompartment. For example, a potato polysaccharide preparation providedherein or a nutritional supplement composition provided herein can beused to increase expression of a LRP1 polypeptide, an APBB1 polypeptide,an IDE polypeptide, a GPX3 polypeptide, or a combination thereof. Inhumans, a composition containing a potato polysaccharide preparationprovided herein or a potato polysaccharide preparation provided hereincan be used to increase expression of a human LRP1 polypeptide, a humanAPBB1 polypeptide, a human IDE polypeptide, a human GPX3 polypeptide, ora combination thereof, in human adipocytes.

In some cases, a potato polysaccharide preparation provided herein or anutritional supplement composition provided herein can be used toincrease expression of polypeptides involved with the metabolism ofCNS-derived amyloid beta polypeptides within the white blood cell tissuecompartment. For example, a potato polysaccharide preparation providedherein or a nutritional supplement composition provided herein can beused to increase expression of an APBB1 polypeptide, an IDE polypeptide,a GPX4 polypeptide, or a combination thereof. In humans, a compositioncontaining a potato polysaccharide preparation provided herein or apotato polysaccharide preparation provided herein can be used toincrease expression of a human APBB1 polypeptide, a human IDEpolypeptide, a human GPX4 polypeptide, or a combination thereof, inhuman white blood cells.

In some cases, a potato polysaccharide preparation provided herein or anutritional supplement composition provided herein can be used toincrease or decrease expression of polypeptides involved with oxidativestress and proinflammatory pathways. For example, a compositioncontaining a potato polysaccharide preparation provided herein or apotato polysaccharide preparation provided herein can be used todecrease expression of an IGF1 polypeptide, a NOS2 polypeptide, or acombination thereof. In humans, a composition containing a potatopolysaccharide preparation provided herein or a potato polysaccharidepreparation provided herein can be used to decrease expression of anIGF1 polypeptide, a NOS2 polypeptide, or a combination thereof, in humanwhite blood cells.

The document will provide addition description in the followingexamples, which do not limit the scope of the invention described in theclaims.

EXAMPLES Example 1—Identification and Characterization of a PotatoPolysaccharide Preparation

6 grams of a Russet potato variety of the Solanum tuberosum species werehomogenized with a Polytron homogenizer in 20 mL water in a 50 mLcentrifuge tube and kept at room temperature for 1 hour. The homogenatewas centrifuged at 4000 rpm for 10 minutes. A Sep-Pak Plus C-18cartridge was activated with 10 mL 100% acetonitrile (ACN) and washedwith 10 mL 0.05% trifluoroacetic acid in water (TFA water). 10 mL of thesupernatant was loaded onto the cartridge, and all H₂O that passesthrough cartridge was collected in 1.5 mL Eppendorf tubes. Next, 10 mLof 2% ACN (in 0.05% TFA water) was passed through the column, and theelutriate was collected in 1.5 mL Eppendorf tubes. Next, 10 mL of 5% ACN(in 0.05% TFA water) was used to wash the column, and the elutriate wascollected in 1.5 mL Eppendorf tubes. Finally, 10 mL of 10% ACN (in 0.05%TFA water) was collected in 1.5 mL Eppendorf tubes after passing throughthe column. All of the fractions were dried, and the dried fractions ofthe same ACN concentration were reconstituted into 1 tube in 1 mL of0.05% TFA water for further purification via HPLC or reconstituted in 1mL of phosphate buffered saline for use in cell treatments.

A Waters 2695 separations module with a photodiode array detector wasused to purify the 10% ACN extract. An XterraRP C18 column (4.6×150 mm)was used for the separation with 0.05% TFA water as the mobile phase.Each HPLC run was a 20 minute gradient ranging from 0 to 2.5% ACN. Theinjection volume was 100 μL, and the flow rate was 0.5 mL/minute. HPLCfractionation of the 10% ACN extract yielded three major UV absorbingpeaks eluted at 3.5, 3.9, and 12.1 minutes (FIG. 1). Collection and HPLCre-purification of the 3.5 minute fraction yielded a symmetrical peakdisplaying a maximum absorbance at 198.3 nm (FIG. 2).

Further chemical characterization of the symmetrical 3.5 minute HPLCpeak material was performed. Pooled 3.5 minute HPLC fractions were driedand reconstituted in 1 mL TFA water and subjected to tandem LC/MS/MS(FIG. 3) and NMR chemical analyses (FIGS. 4 and 5). For the NMRanalysis, ¹H-NMR was run on the sample using deuterium oxide (D₂O) as asolvent to further analyze the sample. The water peak at 4.65 PPM wassolvent-suppressed, and the spectrum was acquired for several hours.Acetamide was detected at 3.2 PPM, along with acetonitrile at 1.9 PPM.Minor peaks were detected at 1.05 PPM, 1.17 PPM (broad peak), 1.189 PPM,and 1.864 PPM. One characteristic of polymeric materials in a proton NMRwas the broadening of peaks such as the shift at 1.17 PPM. These shiftson the NMR could represent the peak at 4.8 PPM and suggested a polar,water-soluble polymer such as a polysaccharide. Taken together, theseresults confirmed the presence of high molecular weight polysaccharidematerial contained in HPLC purified fractions eluting at 3.5 minutes.

Further analysis confirmed that the HPLC purified fraction eluting at3.5 minutes contains polysaccharide material (e.g., highly substitutedcomplex xyloglucan material). To make the polysaccharide materialanalyzable by gas chromatography/mass spectroscopy (GC/MS), it wasconverted into its derivatized carbohydrate fragments. Briefly, thesample was concentrated to a dry residue that was hydrolyzed usingtrifluoroacetic acid. This was then reduced using sodium borohydride,and after borate removal, the end product was acylated using aceticanhydride and pyridine. The end products of the reaction were injecteddirectly on GC/MS to identify any acylated carbohydrates. Based on theend analysis, a larger carbohydrate existed in the sample. The total ionchromatogram (TIC) is shown below in FIG. 6 with appropriate peak labelsbelow in Table 1. The major components identified are indicated in bold(peaks 3, 12, 14, and 21). The corresponding fragmentation for eachcompound is provided in FIGS. 7-26. For each fragmentation, the peakfragmentation pattern is on the top, the compound library fragmentationmatch is on the bottom, and an overlay of the two is in the center.Finally, unlabeled peaks were either column bleed or did not have asufficient match to the compound library.

TABLE 1 Summary of GC/MS results. Retention Time Peak (min) CompoundName Structure 1 10.731 Diacetamide

2 13.669 3-Acetoxy pyridine

3 19.568 3,4-Furan dimethanol, diacetate

4 19.950 1,2,3-propanetriol diacetate

5 23.387 Imidazole, 2- acetamino-5-methyl

6 23.499 6,7-dihydro-5H- pyrrol[2,1,c][1,2,4] triazole-3-carboxylic acid

7 24.304 Acetic acid, 1-(2- methyltetrazol-5-yl) ethenyl ester

8 25.538 1,2,3,4-butanetriol, tetraacetate

9 27.412 (1,5)β(1,3)triacetyl D-galactosan (stereoisomer 1)

10 28.188 (1,5)β(1,3)triacetyl D-galactosan (stereoisomer 2)

11 29.210 Pentaerythritol tetraacetate

12 29.727 1,2,3,4,5-penta-o- acetyl-D-xylitol (isomer 1)

13 30.697 1,2,345-penta-o- acetyl-D-xylitol (isomer 2)

14 32.477 3,5-diacetoxy-benzyl alcohol

15 32.677 β-D-glucopyranose, pentaacetate

16 33.012 D-mannitol hexaacetate

17 33.106 β-D-galactopyranose, pentaacetate

18 33.206 Galacticol, hexaacetate

19 33.364 Cyclohexane carboxylic acid, 1,2,45- tetrakis(acetoxy),(1α,3α,4α,5β)-(-)

20 33.582 Muco-inositol, hexaacetate

21 33.006 D-glucitol- hexaacetate

22 34.463 Myo-inositol, hexaacetate

These results demonstrate the presence of sugar monomers that serve asbuilding blocks for a larger carbohydrate. It appeared from thesemultiple lines of analysis that the potato polysaccharide preparation isa highly substituted complex xyloglucan.

Example 2—Sweet Potatoes and Multiple Varieties of Potatoes Exhibit thePresence of Potato Polysaccharide Material

Six grams of potato material from multiple varieties of Solanumtuberosum (Organic yellow, Purple, Idaho Russet, and Yukon Gold) and sixgrams of material from sweet potatoes (Ipomoea batatas) were extractedin 20 mL of water. 10 mL of that water was then loaded onto a sep-pakcartridge, and the cartridge was then eluted with 10 mL of 10% ACN. TheACN was then dried, and the residue was dissolved in 1 mL of water. A100 μL injection of this water was assessed using HPLC.

The HPLC chromatograms demonstrated that the amount of the first peak(at 3.5 minutes at 210 nm) was the same for all five types of potatoestested (FIGS. 27-31).

In another experiment, material was extracted from a boiled Purplepotato and analyzed. The peak at 3.5 minutes was not reduced in theboiled potato (FIG. 32).

The 3.5 minute peak from two pooled fraction collections from IdahoRusset potatoes was collected, dried, and reconstituted in 100 μL ofwater. The material was then injected into the HPLC yielding a singlepeak at 3.5 minutes (FIG. 33). Taken together, these results demonstratethat potatoes within the Solanum tuberosum and Ipomoea batatas speciescontain potato polysaccharide material.

Example 3—Analysis of a Potato Polysaccharide Preparation

A potato polysaccharide preparation was purified using HPLC from 3 g ofpurple potato. The potato polysaccharide peak was eluted at about 5minutes (FIG. 34). This peak was obtained using a differentchromatographic column (10 mm×150 mm) as compared to the column used toobtain the 3.5 minute peak. Since the column was a larger preparativecolumn and the flow rate was 1.5 mL/minute, the elution time of thepotato polysaccharide was 5 minutes.

Example 4—Use of Potato Polysaccharide Preparations to Reduce AmyloidBeta Levels in Zucker Diabetic Fatty Rats

To assess the ability of potato polysaccharide preparations to reduceamyloid beta levels within a mammal having Alzheimer's disease, blood,the livers, and abdominal fat from rats of four groups of experimentalanimals were collected, weighed, and examined as described in thisExample.

Extraction and Purification of a Potato Polysaccharide Preparation

Typically, 6 g of potato were homogenized with a Polytron homogenizer in20 mL water in a 50 mL centrifuge tube and kept at room temperature for1 hour. The homogenate was centrifuged at 4000 rpm for 10 minutes andthe supernatant fraction was reserved. 10 mL of the supernatant fractionwas percolated through a Sep-Pak Plus C-18 cartridge previouslyactivated with 10 mL 100% acetonitrile (ACN) followed by 10 mL 0.05%trifluoroacetic acid in water (TFA water). Following successive low ACNwashes, semi-purified potato polysaccharide preparation was eluted in 10mL 10% ACN in 0.05% TFA water. The eluent fraction was dried andreconstituted in 1 mL 0.05% TFA water for further purification via HPLC.

The reconstituted 10% ACN eluent fraction was subjected to HPLCpurification utilizing a Waters Xterra RP C18 column (4.6×150 mm) andWaters 2695 separations module with a photodiode array detector. HPLCpurification employed a shallow 20 minute gradient ranging from 0 to2.5% in 0.05% TFA water at a flow rate of 0.5 mL/min. Collection andHPLC re-purification of a major 198 nm UV absorbing peak at 3.5 minutesyielded a symmetrical HPLC peak containing highly purified potatopolysaccharide preparation. The purified HPLC fraction was dried andreconstituted in phosphate buffered saline (PBS) for use in biologicalexperiments.

Potato Polysaccharide Preparation Formulation

Purified potato polysaccharide preparation (10 mL stock solution at 5mg/mL concentration) was stored at 4° C. The vehicle for the study wassterile water (Catalog number 002488, Butler Schein). Each week, thestock solution was diluted 1:100 in sterile water (0.05 mg/mL) anddispensed into daily aliquots. All vehicle and drug solutions werestored at 4° C. and administered at room temperature daily by oralgavage (PO) in a volume of 1 mL/animal (0.15 mg/kg dose based onestimated body weight of 350 g).

In Vivo Animal Model

The Zucker Diabetic Fatty (ZDF) rat model was used (Carley and Severson,Biochim. Biophys. Acta, 1734:112-26 (2005)).

Experimental Animals

Twenty-two 7-week old, male Zucker Diabetic Fatty rats (ZDF, Code: 370)and twenty-two 7-8 week old, male ZDF Lean rats (Code: 371) werepurchased from Charles Rivers Laboratories (Wilmington, Mass.). Thestudy animals were allowed an acclimation period of 4 days prior tobaseline blood collections, at which time two extra animals from eachstrain were dropped from the study based on baseline body weight. Therats were housed two per cage and maintained in the Innovive cagingsystem (San Diego, Calif.) upon arrival at PhysioGenix, Inc. Cages weremonitored daily to ensure the Innovive system maintained 80 air changesper hour and positive pressure. In accordance with the Guide for Careand Use of Laboratory Animals (Eighth Edition), rat rooms weremaintained at temperatures of 66-75 degrees Fahrenheit and relativehumidity between 30% and 70%. The rooms were lit by artificial light for12 hours each day (7:00 AM-7:00 PM). Animals had free access to waterand Purina 5008 rodent food (Waldschimdt's, Madison, Wis.) for theduration of the study except during fasted experiments.

Dosing and Grouping

Two types of rats were used for the study: homozygous obese ZDF/ZDF andheterozygous lean littermates. The rats within the groups were thenchosen at random and divided into groups of 10. Group 1 was thehomozygous ZDF/ZDF vehicle fed rats, group 2 was the homozygous ZDF/ZDFpotato polysaccharide preparation fed, group 3 was the lean vehicle fedrat and group 4 was the lean potato polysaccharide preparation fed rats.The vehicle was distilled water and the potato polysaccharidepreparation was given daily each morning via oral gavage at a dosage of0.05 mg per animal. The dose was usually given in 1 mL of water. Ratswere caged in groups and maintained in 12 hours light/12 hours dark (7am-7 pm). The study lasted for 28 days and all animals were euthanizedby isoflurane overdose and thoracotomy following the collection offasted blood glucose data on Day 28 of the Study. Blood was collectedvia descending vena cava. Liver and abdominal fat were collected,weighed, and a portion of the left lateral liver lobe and abdominal fatwere placed into individual histology cassettes and snap frozen inliquid nitrogen. General pathological observations were recorded.

RNA Isolation

Total RNA extracted from rat tissue samples was isolated and purifiedusing the RNeasy mini kit (Qiagen, Valencia, Calif.). Typically, 100 mgof tissue was resuspended in 1.8 mL of RLT lysis buffer (Qiagen) andhomogenized with a polytron homogenizer for 30 seconds. Blood RNA wasisolated using the PAX RNA kit (Qiagen).

DNA Microarray Analyses

DNA microarray analyses were performed using a system provided byAgilent. Arrays included four arrays per chip (Agilent 4×44K chip).Total RNA was reverse transcribed (1000 ng) using T7 primers and labeledand transcribed using Cyanine-3 dye. Each array was hybridized with atleast 1.65 μg of labeled cRNA at 65° C. for 18 hours. Arrays werescanned using an Agilent array scanner. The microarray platform candetermine a minimum of a 15% change in gene expression.

Real-Time PCR Analyses

Real-time PCR analysis of gene expression was performed to validate theDNA microarray data sets. GAPDH was used as a reference gene. Thereal-time PCR master mix included 25 μL 2× universal master mix, 2.5 μL20× detector set (with the primer and probe), and 21.5 μL of water. PCRwas performed in an Applied Biosystems 7500 sequence detection system.The thermocycler conditions included denaturation at 95° C. for 15seconds and annealing/extension at 60° C. for 60 seconds. Forty cyclesof PCR were preceded by 95° C. for 10 minutes. Reactions were performedin triplicate. The relative quantities of genes were determined usingthe formula 2-AACt using the Applied Biosystems 7500 software.

Results

In vivo administration of purified potato polysaccharide preparation toZDF rats (n=10) vs. vehicle control ZDF rats engendered a statisticallysignificant enhancement of interactive LRP1, APBB1, IDE, and GPX3 geneexpression, normalized as fold changes of 1.8, 1.9, 1.5, and 2.4,respectively, in adipose tissue samples, as depicted in Table 2. Insimilar fashion, in vivo administration of purified potatopolysaccharide preparation to ZDF rats (n=7) vs. vehicle control ZDFrats engendered a statistically significant enhancement of interactiveAPBB1, IDE and GPX4 gene expression, normalized as fold changes of 1.7,1.5, and 2.6, respectively, in blood samples, as depicted in Table 3. Invivo administration of purified potato polysaccharide preparation to ZDFrats (n=7) vs. vehicle control ZDF rats engendered a highly dramatic andstatistically significant reduction of IGF1 and NOS2 proinflammatorygene expression, normalized as fold changes of −1.7 and −4.6,respectively, in blood samples, as depicted in Table 4.

TABLE 2 Enhanced gene expression of LRP1, APBB1, IDE, and GPX3 inadipose tissues of Zucker Diabetic Fatty vs. vehicle control ZDF rats (n= 10) following in vivo potato polysaccharide preparationadministration. Gene Fold Symbol Change Description p value LRP1 1.8 Lowdensity lipoprotein receptor-related 0.02 protein 1 APBB1 1.9 Amyloidbeta precursor protein-binding, 0.0004 family B, member 1 IDE 1.5Insulin degrading enzyme 0.01 GPX3 2.4 Glutathione peroxidase 3 0.0004GPX4 −1.1 Glutathione peroxidase 4 0.6 IGF1 ND Insulin like growthfactor — NOS2 ND Nitric oxide synthase 2, inducible — Data sets werederived by DNA microarray analyses, as described above. ND = geneexpression not detected.

TABLE 3 Enhanced gene expression of APBB1, IDE, and GPX4 in bloodsamples of Zucker Diabetic Fatty vs. vehicle control ZDF rats (n = 7)following in vivo potato polysaccharide preparation administration. GeneFold Symbol Change Description p value APBB1 1.7 Amyloid beta precursorprotein-binding, 0.04 family B, member 1 IDE 1.5 Insulin degradingenzyme 0.04 GPX4 2.6 Glutathione peroxidase 4 0.002 GPX3 ND Glutathioneperoxidase 3 — LRP1 ND Low density lipoprotein receptor-related —protein 1 Data sets were derived by DNA microarray analyses, asdescribed above. ND = gene expression not detected.

TABLE 4 Reduced expression of IGF1, and NOS2 proinflammatory genes inblood samples of Zucker Diabetic Fatty vs. vehicle control ZDF rats (n =7) following in vivo potato polysaccharide preparation administration.Gene Fold Symbol Change Description p value IGF1 −1.7 Insulin-likegrowth factor 1 0.02 NOS2 −4.8 Nitric oxide synthase 2, inducible 0.003Data sets were derived by DNA microarray analyses, as described above.Real-time PCR analysis of IDE expression was performed to validate theDNA microarray data sets. In particular, the enhancement of IDE geneexpression in adipose tissue and blood leukocyte samples, as monitoredby DNA microarray analyses, was confirmed by real time PCR analyses. Asdepicted in FIG. 37, the enhancements in IDE gene expression weredetermined to be 44.9±2.8% (n=9, p=0.02) and 51.3±6.2% (n=6, p=0.04),for adipose tissue and blood samples, respectively. The respective realtime PCR traces are depicted in FIGS. 35 and 36.

These results demonstrate that potato polysaccharide preparations can beused as anti-neurodegenerative agents to reduce amyloid beta levelswithin a mammal having Alzheimer's disease.

Example 5—Use of Potato Polysaccharide Preparations to Improve Cognitionof APP SWE/PSEN1dE9 Mice Dosing and Grouping

APP SWE/PSEN1dE9 mice are used as a model for Alzheimer's disease. Themice within the groups are chosen at random and divided into groups of Xor Y. Groups are given dosages of potato polysaccharide preparation at arange of 0 mg/kg/day (control) up to 0.5 mg/kg/day. One dose is 0.15mg/kg/day. The vehicle is distilled water, and the potato polysaccharidepreparation is given daily each morning via oral gavage at the dosage tobe evaluated. The dose is usually given in 1 mL of water. Mice are cagedin groups and maintained in 12 hours light/12 hours dark (7 am-7 pm).The study proceeds for several months. A typical protocol starts withpotato polysaccharide preparation administration at 20 weeks of age andcontinues through 30 weeks of age and includes 3 groups (20animals/group): vehicle, 5 μg/day potato polysaccharide preparation, and20 μg/day potato polysaccharide preparation.

Data Collection

Body weights are recorded weekly. Whole blood, serum, and plasma arecollected at day 0 for baseline analysis. Whole blood, serum, and plasmaare collected at termination. Brain tissue and vascular are collectedand snap frozen in liquid nitrogen at termination. Whole genomemicroarrays are performed with the frozen tissue samples. Whole blood ispreserved in PAX RNA blood tubes for possible gene expression analysis.Histology to determine the amount of plaque development in animal braintissue is performed at termination. Any appropriate method to determineAPP or AP4, amyloid beta polypeptides in blood or tissue is used.Cognition of the study animals is assessed by using the Morris watermaze methodology.

Other Embodiments

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed is:
 1. A method for reducing amyloid beta levels withina mammal having Alzheimer's disease, wherein said method comprisesadministering to said mammal a composition comprising a potatopolysaccharide preparation obtained from raw potatoes, wherein the levelof amyloid beta within said mammal is reduced.
 2. The method of claim 1,wherein said mammal is a human.
 3. The method of claim 1, wherein saidlevel of amyloid beta within said mammal is reduced in blood.
 4. Themethod of claim 1, wherein said level of amyloid beta within said mammalis reduced in adipose tissue.
 5. The method of claim 1, wherein saidcomposition comprises said potato polysaccharide preparation in anamount that results in between 0.05 mg and 50 mg of the potatopolysaccharide component of said potato polysaccharide preparation beingadministered to said mammal per kg of body weight of said mammal.
 6. Themethod of claim 1, wherein said composition comprises between 1 mg and100 mg of said potato polysaccharide preparation.
 7. The method of claim1, wherein said composition comprises between 6 mg and 20 mg of saidpotato polysaccharide preparation.
 8. The method of claim 1, whereinsaid composition comprises between 1 mg and 100 mg of the potatopolysaccharide component of said potato polysaccharide preparation. 9.The method of claim 1, wherein said composition comprises between 6 mgand 20 mg of the potato polysaccharide component of said potatopolysaccharide preparation.
 10. The method of claim 1, wherein saidcomposition is in the form of a tablet.
 11. The method of claim 1,wherein said composition comprises alpha lipoic acid.
 12. The method ofclaim 1, wherein said composition comprises alpha tocopherol.
 13. Themethod of claim 1, wherein said potato polysaccharide preparation is inan amount that results in between 0.075 mg and 0.5 mg of the potatopolysaccharide component of said potato polysaccharide preparation beingadministered to said mammal per kg of body weight of said mammal. 14.The method of claim 1, wherein at least about 80 percent of said potatopolysaccharide preparation is potato polysaccharide.
 15. The method ofclaim 1, wherein at least about 90 percent of said potato polysaccharidepreparation is potato polysaccharide.
 16. The method of claim 1, whereinat least about 95 percent of said potato polysaccharide preparation ispotato polysaccharide.